nmol l tumor necrosis factor  (R&D Systems)

Journal: International Journal of Molecular Medicine

Article Title: Triptolide exerts antiviral effects and alleviates influenza A-induced pneumonia by inhibiting the overactivation of absent in melanoma 2 signaling in immune cells

doi: 10.3892/ijmm.2026.5829

Figure Lengend Snippet: H1N1 infection affects cell viability, inflammatory cytokine secretion and interactions between HBEpiCs and THP-1 cells. (A) CCK-8 assay revealed that HBEpiC viability decreased in a concentration-dependent manner following H1N1 infection. (B) ELISA revealed that the levels of IL-1β, IL-6, TNF-α, and IL-8 in HBEpiCs decreased with increasing H1N1 infection. (C) CCK-8 assay indicated that supernatants from H1N1-infected HBEpiC cultures reduced the viability of THP-1 cells in a dose-dependent manner. (D) ELISA results suggested that the levels of inflammatory cytokines (IL-1β, IL-6, TNF-α and IL-8) in THP-1 cells were decreased following exposure to supernatants from H1N1-infected HBEpiC cultures. (E) Cell adhesion assay revealed that the number of THP-1 cells adhering to HBEpiCs increased with increasing H1N1 concentration (scale bar, 10 μ m). Arrow indicates THP-1 cells that remain attached to the surface of HBEpiCs, highlighting the adhesion interaction between the two cell types. (F) Transwell assay suggested that H1N1 infection enhanced the migration capacity of THP-1 cells, with increased migration observed at higher virus concentrations (scale bar, 50 μ m). The data are presented as the mean ± standard deviation; ** P<0.01, *** P<0.001 vs. control. H1N1, influenza A; HBEpiCs, human bronchial epithelial cells; ELISA, enzyme-linked immunosorbent assay; IL, interleukin; TNF-α, tumor necrosis factor-α; Con, control; MOI, multiplicity of infection.

Article Snippet: Cell supernatants were collected and analyzed using Human TNF-α High Sensitivity ELISA Kit [cat. no. EK182HS; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.], Human IL-8 ELISA Kit [cat. no. EK108; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.], a human IL-1β ELISA kit [EH0185; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.] and IL-6 [cat. no. EK1217; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.] according to the manufacturer's instructions.

Techniques: Infection, CCK-8 Assay, Concentration Assay, Enzyme-linked Immunosorbent Assay, Cell Adhesion Assay, Transwell Assay, Migration, Virus, Standard Deviation, Control

Journal: International Journal of Molecular Medicine

Article Title: Triptolide exerts antiviral effects and alleviates influenza A-induced pneumonia by inhibiting the overactivation of absent in melanoma 2 signaling in immune cells

doi: 10.3892/ijmm.2026.5829

Figure Lengend Snippet: TP modulates the inflammatory response and immune cell activity in H1N1-infected HBEpiCs and THP-1 cells. (A) No significant changes were observed in HBEpiCs treated with various concentrations of TP (5, 10 and 20 nM) following H1N1 infection compared with the control. (B) After TP treatment, the levels of the inflammatory cytokines IL-1β, IL-6, TNF-α and IL-8 in HBEpiCs were markedly lower than those in the untreated group. (C) The viability of THP-1 cells pretreated with H1N1-infected HBEpiC culture supernatant decreased after TP treatment. (D) The levels of IL-1β, IL-6, TNF-α, and IL-8 in THP-1 cells were markedly lower after TP treatment. (E) The adhesion of THP-1 cells to HBEpiCs induced by H1N1 infection decreased in a dose-dependent manner with increasing TP concentration (scale bar, 10 μ m). Arrow indicates THP-1 cells that remain attached to the surface of HBEpiCs, highlighting the adhesion interaction between the two cell types. (F) The migration capacity of THP-1 cells was markedly reduced when the supernatant from H1N1-infected HBEpiC cultures was treated with TP (scale bar, 50 μ m). The data are presented as the mean ± standard deviation; * P<0.05, ** P<0.01, *** P<0.001 vs. control. TP, triptolide; H1N1, influenza A; HBEpiCs, human bronchial epithelial cells; IL, interleukin; TNF-α, tumor necrosis factor-α; Con, control.

Article Snippet: Cell supernatants were collected and analyzed using Human TNF-α High Sensitivity ELISA Kit [cat. no. EK182HS; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.], Human IL-8 ELISA Kit [cat. no. EK108; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.], a human IL-1β ELISA kit [EH0185; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.] and IL-6 [cat. no. EK1217; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.] according to the manufacturer's instructions.

Techniques: Activity Assay, Infection, Control, Concentration Assay, Migration, Standard Deviation

Journal: International Journal of Molecular Medicine

Article Title: Triptolide exerts antiviral effects and alleviates influenza A-induced pneumonia by inhibiting the overactivation of absent in melanoma 2 signaling in immune cells

doi: 10.3892/ijmm.2026.5829

Figure Lengend Snippet: AIM2 overexpression reverses the immunosuppressive effects of TP in THP-1 cells. (A) Transfection with Ad-AIM2 upregulated the expression of AIM2 compared with that of Ad-NC in THP-1 cells. (B) AIM2 protein levels were elevated in THP-1 cells overexpressing AIM2 compared with control THP-1 cells. (C) In THP-1 cells, AIM2 overexpression increased the LDH leakage rate. (D) AIM2 overexpression reversed the TP-induced reduction in adhesion between THP-1 cells and HBEpiCs (scale bar, 10 μ m). (E) AIM2 overexpression enhanced the migration ability of THP-1 cells compared with that of H1N1-treated control cells and reversed the TP-induced decrease in THP-1 cell migration (scale bar, 50 μ m). (F) AIM2 overexpression reversed the TP-induced reduction in inflammatory cytokine levels. The data are presented as the mean ± standard deviation; * P<0.05, ** P<0.01, *** P<0.001 vs. Con; # P<0.05, ## P<0.01, ### P<0.001 vs. Ad-AIM2. AIM2, absent in melanoma 2; TP, triptolide; LDH, lactate dehydrogenase; HBEpiCs, human bronchial epithelial cells; H1N1, influenza A; GSDMD, gasdermin D; IL, interleukin; TNF-α, tumor necrosis factor-α; Con, control.

Article Snippet: Cell supernatants were collected and analyzed using Human TNF-α High Sensitivity ELISA Kit [cat. no. EK182HS; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.], Human IL-8 ELISA Kit [cat. no. EK108; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.], a human IL-1β ELISA kit [EH0185; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.] and IL-6 [cat. no. EK1217; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.] according to the manufacturer's instructions.

Techniques: Over Expression, Transfection, Expressing, Control, Migration, Standard Deviation

Journal: JID Innovations

Article Title: Development of a noninvasive device and method for measuring immune parameters in skin disease

doi: 10.1016/j.xjidi.2026.100456

Figure Lengend Snippet: Detection of inflammatory cytokines in skin wash of healthy volunteers and patients with AD and psoriasis. Cytokine concentrations were measured in skin wash fluid using Luminex assay. Inflammatory cytokines ( a ) IL-1β, ( b ) TNFα, ( c ) IL-18, and ( d ) IL-8. AD-related immune mediators ( e ) CCL13, ( f ) CCL18, ( g ) sIL-2R, and ( h ) IL-4. Psoriasis-associated immune mediators ( i ) IL-17A, ( j ) IL-23, ( k ) sTNFR1, and ( l ) IFNγ. Concentrations are depicted as mean ± SD, with individual data points shown. PS denotes psoriasis, NL denotes nonlesional, and L denotes lesional. ∗ P < .05, ∗∗ P < .01, and ∗∗∗ P < .001. AD, atopic dermatitis; HC, healthy control; sIL-2R, soluble IL-2R; sTNFR1, soluble TNFR1.

Article Snippet: E coli –derived recombinant human TNFα, IL-1β, IL-6, IL-8, IL-10, and IL-18 (endotoxin levels <0.10 EU/μg by the limulus amebocyte lysate method) were purchased from R&D Systems and reconstituted in PBS.

Techniques: Luminex, Control

Journal: Redox Report : Communications in Free Radical Research

Article Title: Bilirubin reduces mortality in sepsis models by inhibiting NOX2-mediated formation of neutrophil extracellular traps

doi: 10.1080/13510002.2026.2664962

Figure Lengend Snippet: Effects of bilirubin against CLP-induced sepsis in mice. (A) Mice were subjected to CLP surgery and intravenously injected with bilirubin (40 mg/kg) or vehicle. Mice were monitored at 12-h intervals to check the survival rates. (B) After CLP surgery, all mice received intraperitoneal injection of an antibiotics mixture (ceftriaxone 50 mg/kg and metronidazole 35 mg/kg). Mice were then intravenously administered either bilirubin (40 mg/kg) or vehicle. The state of survival was monitored for 96 h. The survival rates between the bilirubin and the vehicle groups were statistically analyzed using the Mantel-Cox test ( N = 15). (C, D) Biomarkers for NETosis were measured in the plasma of sham or CLP mice treated with vehicle or bilirubin 12 h after injection. Cell-free DNAs were stained with PicoGreen (C) and the H3cit levels were assessed using the ELISA kit (D). (E, F) Pro-inflammatory cytokines TNF-α (E) and IFN-γ (F) were analyzed in the plasma of the mice using specific ELISA kits. Each bar represents the mean ± SEM ( N = 5–8). Asterisks and numbers represent statistical significance and p- values by Mann–Whitney U test, respectively.

Article Snippet: Biochemical analyses were performed using the Human IFN-γ Quantikine ELISA Kit (R&D Systems; Minneapolis, MN), the Human TNF-α Quantikine ELISA Kit (R&D Systems; Minneapolis, MN), the Citrullinated Histone H3 ELISA Kit (Cayman Chemical; Ann Arbor, MI), and the Quant-iTTM PicoGreenTM dsDNA Assay Kit (Invitrogen; Waltham, MA).

Techniques: Injection, Clinical Proteomics, Staining, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: Redox Report : Communications in Free Radical Research

Article Title: Bilirubin reduces mortality in sepsis models by inhibiting NOX2-mediated formation of neutrophil extracellular traps

doi: 10.1080/13510002.2026.2664962

Figure Lengend Snippet: Effects of bilirubin against LPS-induced sepsis in mice. (A) Mice were intraperitoneally injected with 40 mg/kg of LPS and intravenously injected with bilirubin (40 mg/kg) or vehicle. Mice were monitored at 12-h interval to check the survival rates. The survival rates were statistically analyzed using the Mantel-Cox test ( N = 15). (B, C) NETosis markers were measured in the plasma of PBS control or LPS-treated mice 12 h after bilirubin or vehicle injection. The plasma levels of cell-free DNAs and H3cit were analyzed using PicoGreen (B) and the ELISA kit (C), respectively. (D, E) Pro-inflammatory cytokines TNF-α (D) and IFN-γ (E) were analyzed in the plasma using specific ELISA kits. Each bar represents the mean ± SEM ( N = 5–8). Asterisks and numbers represent statistical significance and p- values by Mann–Whitney U test, respectively.

Article Snippet: Biochemical analyses were performed using the Human IFN-γ Quantikine ELISA Kit (R&D Systems; Minneapolis, MN), the Human TNF-α Quantikine ELISA Kit (R&D Systems; Minneapolis, MN), the Citrullinated Histone H3 ELISA Kit (Cayman Chemical; Ann Arbor, MI), and the Quant-iTTM PicoGreenTM dsDNA Assay Kit (Invitrogen; Waltham, MA).

Techniques: Injection, Clinical Proteomics, Control, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: Scientific Reports

Article Title: Plant extract mixture shows anti-inflammatory and barrier-strengthening effects and activates aryl hydrocarbon receptor in a 2D psoriasis model

doi: 10.1038/s41598-026-50000-8

Figure Lengend Snippet: Plant extract mixture downregulates inflammatory markers in the 2D psoriasis model. CaCl 2 -differentiated NHEKs were stimulated as described in Fig. . Gene expression levels of ( a ) IL1A , ( b ) IL1B, ( c ) IL1RN , ( d ) CXCL8 , ( e ) TNFA , ( f ) IL17C , ( g ) IL36G , ( h ) CSF2 , ( i ) VEGFA were measured. Statistical significance was tested by a, b, c, e, h) one-way ANOVA with subsequent Sidak’s multiple comparison test or d, f, g) Kruskal-Wallis test with subsequent Dunn’s multiple comparisons test ( n = 12, * p < 0.05; ** p < 0.01; *** p < 0.001; ns = not significant). Protein expression levels of ( j ) IL-8 and k) TNFα were measured in the supernatants of the cells by ELISA. Values for unstimulated control and plant extract-treated cells were below the detection limit of 31,3 pg/mL (IL-8) or 62,5 pg/mL (TNFα) and not detectable; therefore, no statistical analysis was performed. n.d.= non-detectable.

Article Snippet: TNFα and IL-8 concentrations were determined using commercial ELISA kits (human TNFα or human CXCL8/IL-8 DuoSet ELISA, R&D Systems, Minneapolis, MN, USA) following the manufacturer’s protocol.

Techniques: Plant Extract, Gene Expression, Comparison, Expressing, Enzyme-linked Immunosorbent Assay, Control